Recombinant bacterial RNA polymerase: preparation and applications.
Availability of DNA-dependent RNA polymerase from various bacteria is a key for setting up specific in vitro transcription systems necessary for understanding species-specific transcription regulation. We describe here two main strategies for recombinant RNA polymerase preparation-through in vitro reconstitution and heterologous co-overproduction in Escherichia coli. Both strategies can be used for preparation of large amounts of RNA polymerases from any bacteria for which sequences of rpo (RNA polymerase) genes are known.
Kuznedelov
Konstantin
K
Waksman Institute, Rutgers, The State University of New Jersey, 190 Frelinghuysen Road, Piscataway, NJ 08854, USA. kuznedelo@waksman.rutgers.edu
Severinov
Konstantin
K
D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001419:Bacteria / Q000201:enzymology / Q000235:genetics; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000235:genetics; D012321:DNA-Directed RNA Polymerases / Q000096:biosynthesis* / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D005822:Genetic Vectors / Q000235:genetics; D017510:Protein Folding; D021122:Protein Subunits / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D012808:Sigma Factor / Q000096:biosynthesis / Q000235:genetics; D014158:Transcription, Genetic / Q000187:drug effects; D014170:Transformation, Genetic
D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D021122:Protein Subunits; D011994:Recombinant Proteins; D012808:Sigma Factor; D012321:DNA-Directed RNA Polymerases